(C07D 477/00, A61K 31/40)
The invention belongs to the field of pharmaceutical industry and relates to novel ethylidene derivatives of tricyclic carbapenems in the form of pure diastereoisomers and in the form of pure geometric isomers, to processes for the preparation thereof, to pharmaceutical compositions containing them and to the use thereof in human and veterinary medicine. Novel ethylidene derivatives of tricyclic carbapenems are used as inhibitors of action of xcex2-lactamases and/or as antibiotics.
Due to the phenomenon of resistance to various antibiotics and synthetic chemotherapeutics, there exists a constant need to prepare novel effective antiinfection agents. There also exists a constant need to prepare novel inhibitors of enzymes xcex2-lactamases which are responsible for the resistence against xcex2-lactam antibiotics. Novel ethylidene derivatives of tricyclic carbapenems are chemical compounds which are inhibitors of enzymes xcex2-lactamases and/or antibiotics.
In prior art carbapenem compounds are known as effective antibacterial agents. A novel sub-group of carbapenem compounds are tricyclic carbapenems (first disclosed in EP-A-416953 and EP-A-422596) which are novel antibacterial agents in a broad spectrum of pathogenic gram-negative and gram-positive bacteria as inhibitors of the enzyme D,D-peptidase (A. Perboni et al. in Recent Advances in the Chemistry of Antiinfective Agents, Bentley, H. H., Ponsford, R. Eds., The Royal Society of Chemistry, T. Graham House, Cambridge, 1992, p. 21 and S. Hannesian et al., Bioorg. Med. Chem. Lett. 5, (1995) 2535).
Tricyclic carbapenem compounds with an antibacterial action are also disclosed in U.S. Pat. No. 5,459,260, EP-A-502464, EP-A-502465, EP-A-502468, EP-A-517065, PCT-WO-94/05666, U.S. Pat. Nos. 5,372,993, 5,374,630, PCT-WO-94/21637, PCT-WO-94/21638, PCT-WO-95/03700, PCT-WO-95/13278, PCT-WO-95/23149, JP 06,166688 and JP 08,53459.
The compound 4S,8S-4methoxy-(9R)10S,12R)-10-(1-hydroxyethyl)-11-oxo-1-azatricyclo-[7.2.0.03.8]undec-2-ene (GV104326 or sanfetrinem) is a biologically active molecule with an action on the enzyme D,D-peptidase which is essential for the formation of the cell wall in the process of bacterial multiplication (E. di Mvodugno et al., Antimicrob. Agents Chemother. 38 (1994) 2362).
Bycyclic carbapenem compounds with an antibacterial action are described in EP-A-0005349 and U.S. Pat. No. 4,235,922. There are no data about said compounds being inhibitors of enzyme xcex2-lactamase.
In all hitherto disclosed tricyclic carbapenem compounds there are no data on any inhibition action to enzymes of the xcex2-lactamase species. Thus, the object of the present invention are novel ethylidene derivatives of tricyclic carbapenem compounds with a completely novel structure and with the property of inhibiting the enzyme xcex2-lactamase. This property does not reduce the possibility of potential antibacterial activity thereof.
The invention relates to novel ethylidene derivatives of tricyclic carbapenems of the formula I 
wherein the ring marked C fused to the basic carbapenem nucleus in 3and 4 positions is a five-, six- or seven-membered ring whereat
1. one or more carbon atoms in the ring marked C may be mono or disubstituted with substituents which may be the same or different and may mean:
a) a hydrogen atom.
b) a saturated alkyl chain with 1 to 20 carbon atoms and the saturated alkyl chain may be straight (such as methyl, ethyl, n-propyl, n-butyl) or branched in any position (such as isopropyl, s-butyl, isobutyl, isoamyl, tert-butyl) and each chain member may be mono or disubstituted with substituents such as halo (such as fluoromethyl, trifluoromethyl, 2-chloroethyl), hydroxy (such as hydroxymethyl, 2-hydroxyethyl), (C1-C4)-alkyloxy (such as methoxymethyl, 2-methoxyethyl), mercapto and (C1-C4)-alkylmercapto (such as mercaptomethyl, 2-methylmercaptoethyl), (C1-C4)-alkanesulfonyl (such as methanesulfonylmethyl), amino, (C1-C4)-alkylamino and di-(C1-C4)-alkylamino (such as 2-aminoethyl, 2-methylaminoethyl, 2-dimethylaminoethyl), alkyleneamino (such as 2-(1-piperidinyl)ethyl, 1-pyrrolidinylmethyl), guanidino (such as guanidinomethyl), unsubstituted N1-mono, N3-mono, N1,N3-di and N3,N3-di-(C1-C4)-formamidino (such as iminomethylaminomethyl, 2-(dimethylaminomethyleneamino)ethyl), aromatic or heteroaromatic five- or six-membered ring (such as phenyl, furyl, 2-pyridyl), (C1-C4)-alkyloxycarbonyl (such as carbethoxymethyl), cyano (such as 2-cyanoethyl), oxo (such as acetyl, propionyl, 2-oxopropyl),
c) an unsaturated alkyl chain with 1 to 20 carbon atoms and the unsaturated alkyl chain may be straight with double bonds or triple bonds (such as vinyl, propenyl, allyl, ethinyl, propargyl) or branched in any position with double bonds or triple bonds (such as 2-propenyl) and each chain member may be mono or disubstituted with substituents such as halo, hydroxy, (C1-C4)-alkyloxy, thio and (C1-C4)-alkylthio, (C1-C4)-alkanesulfonyl, amino, (C1-C4)-alkylamino and di-(C1-C4)-alkylamino, aromatic or heteroaromatic five- or six-membered ring (such as phenyl, furyl, 2-pyridyl), (C1-C4)-alkyloxycarbonyl, cyano, oxo,
d) a saturated or partly unsaturated cycloalkyl radical with 3 to 7 members (such as radicals from cyclopropyl to cycloheptyl, cyclohex-1-enyl) and the ring may comprise one or more oxygen, sulfur or nitrogen atoms (such as 2-tetrahydrofuranyl, 1-piperidinyl, 1-pyrrolidinyl) and each ring member may be mono or disubstituted with substituents such as halo, hydroxy, (C1-C4)-alkyloxy, thio and (C1-C4)-alkylthio), (C1-C4)-alkanesulfonyl, amino, (C1-C4)-alkylamino and di-(C1-C4)-alkylamino, (C1-C4)-alkyloxycarbonyl, cyano, oxo,
e) an aromatic or heteroaromatic five- or six-membered ring (such as phenyl, furyl, 2-pyridyl),
f) a hydroxy, C1-C10)-alkyloxy (such as methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, cyclohexyloxy), mono or disubstituted (C1-C10)-alkyloxy (such as 2-hydroxyethyloxy), 2,3-dihydroxyprop-1-yloxy, 2-methoxyethyloxy, fluoromethyloxy, aminoethyloxy, 2-dimethylaminoethyloxy), acyloxy (such as formyloxy, acetyloxy, benzoyloxy, ethoxycarbonyloxy, allyloxycarbonyloxy), mono, di or tri-(C1-C4)-alkylsilyloxy (such as trimethylsilyloxy, tert-butyldimethylsilyloxy) group,
g) a mercapto, (C1-C10)-alkylmercapto (such as methylmercapto, ethylmercapto), mono or disubstituted (C1-C10)-alkylmercapto (such as 2-hydroxyethylmercapto, 2,3-dihydroxyprop-1-ylmercapto, 2-methoxyethylmercapto, 2-mercaptoethylmercapto, 2-methylmercaptoethylmercapto, 2-aminoethylmercapto, 2-dimethylaminoethylmercapto), acylmercapto (such as acetylmercapto) group,
h) an amino, (C1-C4)-alkylamino and di-(C1-C4)-alkylamino (such as methylamino, dimethylamino, 2-hydroxyethylamino, bis(2-hydroxyethyl)amino, aminoethyloxy, 2-dimethylaminoethyloxy, 2-aminoethylamino, 2-piperidinoethylamino), acetylamino, allyloxycarbonylamino, iminomethylamino, N-methylaminomethyleneamino, N,N-dimethylaminomethyleneamino, guanidino, cyanoguanidino, methylguanidino group,
i) a halo atom (such as fluoro, chloro, bromo, iodo),
j) an azido, nitro, cyano, (C1-C4)-alkyloxycarbonyl (such as carbomethoxy, carbethoxy) group,
k) a (C1-C4)-alkanesulfonyl group (such as methanesulfonyl, ethanesulfonyl);
2. one or more carbon atoms in the ring marked C may be substituted with a substituted or unsubstituted alkyl chain which is linked to the ring marked C via double bond in the form of  greater than C*=CR1R2, and C* means a carbon atom in the ring marked C,=means an exocyclic double bond and the substituents R1 and R2, which may be the same or different, may mean:
a) a hydrogen atom so that the substituent is methylene,
b) an unsubstituted saturated alkyl chain with 1 to 20 carbon atoms and the unsubstituted saturated alkyl chain may be straight (such as ethylidene, n-propylidene, n-butylidene, isopropylidene) or branched in any position (such as 2,2-dimethylpropylidene),
c) an unsubstituted unsaturated alkyl chain with 1 to 20 carbon atoms and the unsubstituted unsaturated alkyl chain may be straight or branched with double bonds or triple bonds (such as vinylidene, allylidene),
d) an unsubstituted saturated or partly unsaturated cycloalkyl or heteroaryl with 3 to 7 members (as comprised in groups from cyclopropylmethylene to cycloheptylmethylene),
e) an aromatic or heteroaromatic five- or six-membered ring (as comprised in groups such as benzylidene, diphenylmethylene),
f) a substituted saturated or partly unsaturated alkyl chain or substituted three- to seven-membered carbocyclic ring whereat any carbon atom in the chain or in the ring may be mono or disubstituted with substituents such as halo, hydroxy, (C1-C4)-alkyloxy, (C1-C4)-alkylthio, (C1-C4)-alkanesulfonyl, amino, (C1-C4)-alkylamino, di-(C1-C4)-alkylamino, (C1-C4)-alkyloxycarbonyl, cyano, oxo as comprised in groups such as hydroxyethylidene, methoxyethylidene,
g) a hydroxy, (C1-C4)-alkyloxy, acyloxy, mercapto, (C1-C4)-alkylmercapto, amino, (C1-C4)-alkylamino, di-(C1-C4)-alkylamino and acylamino group as comprised in groups such as hydroxymethylene, methoxymethylene, dimethoxymethylene, dimethylaminomethylene, acetylaminomethylene groups,
h) a nitro, cyano, (C1-C4)-alkyloxycarbonyl group so that the substituent is dicyanomethylene, bis(carbomethoxy)methylene, carbethoxymethylene;
and the substituents R1 and R2 may also mean a joint alkylene chain (CH2)n (n=2 to 7) closed to a ring and any methylene (xe2x80x94CH2xe2x80x94) member may be replaced by oxa (xe2x80x94Oxe2x80x94), thia (xe2x80x94Sxe2x80x94), imino (xe2x80x94NHxe2x80x94) or (C1-C4)-alkylimino group as comprised in groups such as cyclopentylidene, cyclohexylidene, 2-(1,3-dioxacyclopentylidene);
3. one or more carbon atoms in the ring marked C may be substituted with a hetero atom via double bond (such as oxo, thioxo, hydroxyimino, (C1-C4)-alkylimino, acylimino);
4. one or more carbon atoms in the ring marked C may be disubstituted with substituents closed to a ring to obtain a spiro compound and, besides a carbon atom, the ring members may also be oxygen, sulfur and nitrogen atoms (such as ethylene, 1,3-propylene, 1,5-pentylene, ethylenedioxy);
5. one or more carbon atoms in the ring marked C may be replaced by an oxygen atom;
6. one or more carbon atoms in the ring marked C may be replaced with a sulfur atom which may be mono or dioxidized to obtain sulfoxides or sulfones;
7. one or more carbon atoms in the ring marked C may be replaced with a nitrogen atom which may be substituted in the form of  greater than N*-R3 and N* means a nitrogen atom in the ring marked C and R3 may mean:
a) a saturated alkyl chain with 1 to 20 carbon atoms and the unsubstituted saturated chain may be straight (such as methyl, ethyl, n-propyl, n-butyl) or branched in any position (such as isopropyl, s-butyl, isobutyl, isoamyl, tert-butyl) and each chain member may be once or several times substituted with substituents such as halo (such as fluoromethyl, trifluoromethyl, 2-chloroethyl), hydroxy (such as 2-hydroxyethyl), (C1-C4)-alkyloxy (such as 2-methoxyethyl), (C1-C4)-alkylthio (such as methylmercaptoethyl), (C1-C4)-alkanesulfonyl (such as methanesulfonylmethyl), amino, (C1-C4)-alkylamino, di-(C1-C4)-alkylamino (such as 2-aminoethyl, 2-methylaminoethyl, 2-dimethylaminoethyl), aromatic or heteroaromatic five- or six-membered ring (such as phenyl, furyl, 2-pyridyl), (C1-C4)-alkyloxycarbonyl (such as carbethoxymethyl), cyano (such as cyanoethyl), oxo (such as acetyl, propionyl, 2-oxopropyl), imino (such as iminomethyl, aminoiminomethyl, aminocyanoiminomethyl),
b) an unsubstituted unsaturated alkyl chain with 1 to 20 carbon atoms and this unsubstituted unsaturated alkyl chain may be straight with double bonds (such as vinyl, propenyl, allyl) or triple bonds (such as ethinyl, propargyl) or branched in any position with double or triple bonds (such as 2-propenyl),
c) an unsubstituted saturated or partly unsaturated cycloalkyl or heteroaryl radical with 3 to 7 members (such as radicals from cyclopropyl to cycloheptyl, cyclohex-1-enyl, 4-piperidinyl),
d) an unsubstituted aromatic or heteroaromatic five- or six-membered ring (such as phenyl, furyl, 2-pyridyl),
e) groups such as cyano, (C1-C4)-alkyloxycarbonyl (such as carbomethoxy, carboethoxy), aminocarbonyl, (C1-C4)-alkylaminocarbonyl, (C1-C4)-alkylsulfonyl (methanesulfonyl);
and wherein X may mean:
1. a hydrogen atom so that the compound of the formula I is a carboxylic acid; in the case of a basic centre in the molecule, a hydrogen atom is linked to it as a proton, the carboxyl group is in the anion form as carboxylate and the compound of the formula I is in the form of a zwitter ion,
2. an alkali metal so that the compound of the formula I is an alkali metal salt (such as lithium carboxylate, sodium carboxylate, potassium carboxylate),
3. an earth alkali metal so that the compound of the formula I is an earth alkali metal salt wherein for one bivalent metal ion there are two carboxylate anions (such as calcium dicarboxylate),
4. the ammonium ion or a protonated form of mono, di or trisubstituted acyclic or cyclic aliphatic amine or a protonated form of some other nitrogen base. The compound of the formula I is in this case a salt of a carboxylic acid and ammonia or amine (such as trimethylamine, triethylamine, N,Nxe2x80x2-dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, 2-piperidinyl) or amidine (such as 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene, 3,3,6,9,9-pentadimethyl-2,10-diaza-bicyclo[4.4.0]dec-1-ene) or guanidine (such as guanidine, cyanoguanidine) or some other nitrogen base (such as 4-dimethylaminopyridine, imidazole),
5. the quaternized ammonium ion so that the compound of the formula I is a corresponding quaternary ammonium carboxylate (such as tetrabutylammonium carboxylate),
6. a radical R4 whereat the compound of the formula I is in the ester form and the radical R4 may be:
a) selected from the group comprising (C1-C20)-alkyl (such as methylethyl, tert-butyl), (C1-C20)-alkenyl (such as allyl), substituted alkyl (such as (C1-C4)-alkoxyalkyl, (C1-C4)-alkylthioalkyl, phenetyl, 2,2,2-trichloroethyl, 2-oxo-5-methyl-1,3-dioxolene-4-yl)methyl, benzyl, p-methoxybenzyl, p-nitrobenzyl, o-nitrobenzyl, bis(methoxyphenyl)-methyl, 3,4-dimethoxybenzyl, benzhydryl, trityl, 2-trimethylsilylethyl), substituted silyl (such as trimethylsilyl, tert-butyldimethylsilyl), phthalidyl etc.,
b) a radical which may be presented in a following form 
xe2x80x83wherein
R5 represents hydrogen or a lower alkyl with 1 to 4 carbon atoms,
R6 represents hydrogen, alkyl, cycloalkyl, alkoxy, cycloalkoxy, cycloalkylalkyl, alkenyloxy, phenyl,
so that the compounds of the formula I are biologically degradable esters, which are known from the group of cefalosporin antibiotics as prodrug agents (such as 1-pivaloyloxymethyl, 1-pivaloyloxyethyl, acetoxyethyl, 1-acetoxyethyl, 1-methoxy-methylethylcarbonyloxymethyl, 1-(1-methoxy-1-methylethylcarbonyloxy)ethyl, 1-benzoyloxyethyl, 1-(isopropoxycarbonyloxy)-ethyl, cyclohexyloxy-carbonyloxymethyl esters).
The invention relates to novel ethylene derivatives of tricyclic carbapenems of the general formula I in the form of pure diastereoisomers and in the form of pure geometric isomers. The compounds of the formula I comprise at least 2 pure geometric isomers when the methyl group in the ethylidene substituent in 6 position of the carbapenem ring of the compound of the formula I is configured around a double bond as (Z) or as (E) and at least 2 pure diastereoisomers since a new chiral centre in 4 position, which is formed in a joint point with the new ring, may be configured as (R) or as (S). In Scheme 1 and in all further schemes the bold bond represents the position above the level of the sheet and the broken line represents the position under the level of the sheet. The mark (R) or (S) depends on the kind of ring marked C and on the substituents bound to the ring marked C and is determined according to Cahn-Ingold-Prelog rule (Cahn et al., Experientia 12, (1956) 81).
The configuration in 5 position in the joint point of the four- and five-ring of the compound of the general formula I is always the same and is always under the level of the sheet and the mark (R) or (S) is determined according to the above-mentioned Cahn-Ingold-Prelog rule.
Scheme 1: Novel ethylidene derivatives of tricyclic carbapenems in the form of pure geometric isomers and in the form of pure diasteroisomers 
The following compounds of the formula I are especially important as inhibitors of the action of enzymes xcex2-lactamases and/or as antibiotics:
(8S,9R)-10-((E)-ethylidene)-11-oxo-1-azatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylic acid, a pharmaceutically acceptable salt or ester thereof,
(8R,9R)-10-((E)-ethylidene)-11-oxo-1-azatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylic acid, a pharmaceutically acceptable salt or ester thereof,
(8S,9R)-10-((E)-ethylidene)-11-oxo-1-aza-6-thiatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylic acid, a pharmaceutically acceptable salt or ester thereof,
(8R,9R)-10-((E)-ethylidene)-11-oxo-1aza-6-thiatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylic acid, a pharmaceutically acceptable salt or ester thereof.
The compounds of the formula I in the form of an acid and of pharmaceutically acceptable salts or esters thereof are used as antibacterial agents having bacteriostatic and bactericide action upon gram-positive and/or gram-negative bacteria or upon other infects and are used in the prevention and treatment of infectious diseases in humans and animals. The object of the invention is thus also the use of compounds of formula I and of pharmaceutically acceptable salts or esters thereof in the treatment of bacterial diseases in humans and animals, which diseases are caused by gram-positive and/or gram-negative bacteria and other microorganisms.
The compounds of the formula I in the form of an acid and of pharmaceutically acceptable salts or esters thereof are used also and especially as inhibitors of enzymes xcex2-lactamases in a pharmaceutical formulation or in a pharmaceutical formulation in combination with other xcex2-lactam antibiotics such as penicillins and cefalosporins. Thus there may be achieved an effective antibacterial action also upon microorganisms resistent to xcex2-lactam antibiotics. The mechanism of action of enzymes xcex2-lactamases is already well investigated (J. Knowles, Acc. Chem. Res. 18, (1985) 97) and may be described in short as a hydrolysis of the amide bond of xcex2-lactam antibiotic thus losing its ability to inhibit the enzyme D,D-peptidase, this ability being a condition for the antibiotic action of a xcex2-lactam antibiotic. The process of inhibition of the enzyme xcex2-lactamase takes place in two steps. The first step is acylation which generates a tetrahedron complex between a xcex2-lactam inhibitor in Ser70 site in the protein sequence, and this causes a blocade of the active site of the enzyme responsible for the action upon an antibiotic. The second step is deacylation which takes place via action of water and amino acid residues of the active site of the enzyme xcex2-lactamase upon a covalently bound inhibitor-enzyme complex. The whole inhibition process is similar to a process wherein the same enzyme decomposes xcex2-lactam antibiotics (essential differences existing only in the kinetics of the first and the second process steps) and thus the structures of xcex2-lactam antibiotics and inhibitors of the enzyme xcex2-lactamase are similar.
It is characteristic for antibiotics of penam, carbapenem, cefem and carbacefem types that they are substituted with acylamino groups or with a 1-hydroxyethyl group in 6 positions (penams) and 7 positions (cefems), yet these compounds show a relatively poor inhibitory action upon the enzyme ≢-lactamase. Also for such compounds from the field of tricyclic carbapenems (as disclosed in EP-A-0422596) only an antibiotic action has been known so far. It has been shown that the 6 position in the inhibitors of the enzyme xcex2-lactamase must be differently substituted. 6-Ethylidene substituted carbapenems were found to be available systems since they may be obtained from intermediates that are also used in the synthesis of carbapenem antibiotics. Although no compound among them has hitherto been known to act as an inhibitor of the enzyme xcex2-lactamase, it has been found by means of the method of molecular modeling i.e. by calculations and comparison of electronic densities of inhibitors of the enzymes xcex2-lactamases in clinical use and of substrates of xcex2-lactamases of penem and cefem types (D. Kocjan and T. {haeck over (S)}olmajer, QSAR and Molecular Modeling, Ed. F. Senz, J. Ginaldo and F. Monant, Computational Tools and Biological Applications, Prous Science Publishers, Barcelona 1995, pp. 335-337) that the calculated distribution of electronic density of the compounds of the formula I and of pharmaceutically acceptable salts or esters thereof could be suitable for inhibitory action upon the enzyme xcex2-lactamase. It has surprisingly been found that the compounds of the formula I according to the invention are active as inhibitors of the enzyme xcex2-lactamase and thereby they differ from the known carbapenem antibiotics disclosed in the literature. The results of testing the inhibitory action of the compounds of the formula I also confirm their inhibitory action upon the enzyme xcex2-lactamase.
Determination of inhibitory action of the compounds of the formula I upon the enzyme xcex2-lactamase
The inhibitory action of the compounds of the formula I was determined according to standard methodology (Bush, K. and Sykes, R. B. Methods of Enzymatic Analysis, Third Edition, Vol. 4, p. 280, Verlag Chemie GmbH, Weinheim, 1984). The method comprises a spectrophotometric quantitative (band at 495 nm) measurement of catalytic action of the enzyme xcex2-lactamase from the E. coli EC 3.5.2.6 strain. As a substrate nitrocefin ((7R)-3-((E)-2,4-dinitrostiryl)-7-(2-thienylacetamido)-3-cefem-4-carboxylic acid) was used which has the broadest spectrum of susceptibility and sensibility. The hydrolysis of nitrocefin in the presence of an inhibitor of the enzyme xcex2-lactamase was determined.
Hydrolysis of nitrocefin with xcex2-lactamase of Escherichia coli 3.5.2.6 in the presence of inhibitors
Experimental Data:
Legend:
/no inhibitor
compound A sodium (8R,9R)-10-((E)-ethylidene)-11-oxo-1-aza-6-thiatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylate
compound B sodium (8S,9R)-10-((E)-ethylidene)-11-oxo-1-azatricyclo[7.2.0.03,8]undec-2-ene-2-carboxylate
The object of the invention is thus also the use of the compounds of the formula I, pharmaceutically acceptable salts or esters thereof as inhibitors of the enzyme xcex2-lactamase in human and veterinary medicine.
The invention also relates to pharmaceutical formulations for the treatment of infections in humans and animals, which formulations contain as an active ingredient a therapeutically active amount of a compound of the formula I in the form of an acid or of pharmaceutically acceptable salt or ester thereof, optionally in a combination with a xcex2-lactam antibiotic, together with pharmaceutically acceptable carriers and other pharmaceutically acceptable auxiliary substances for peroral, parenteral, rectal, topical, ophthalmological, nasal or genito-urinary applications in human and veterinary medicine.
Parenteral pharmaceutical formulations according to the invention are stored in ampoules in the form of suspensions, emulsions or solutions in oily carriers or in water together with auxiliary substances such as suspending agents, stabilizers, dispersants and/or preservatives, or in containers with a lid, optionally together with added preservative in the form of a powder, and the contents are dissolved before use in a solvent, most frequently in pyrogen-free water. Parenteral pharmaceutical preparations according to the invention may be applied intravenously or intramuscularly.
Pharmaceutical formulations according to the invention may be used perorally in the form of solid formulations such as tablets, capsules, pastilles, pills, granules, powders together with binders such as syrup, arabic gum, gelatine, sorbitol, tragacanth and polyvinylpyrrolidone, with fillers such as lactose, saccharose, corn starch, calcium phosphate, sorbitol, glycine, with lubricants such as magnesium stearate, talc, polyethylene glycol, silica, with disintegrants such as potato starch, and wetting agents such as sodium lauryl sulfate. Tablets and granules may be also coated with a suitable film according to known processes. Peroral pharmaceutical formulations according to the invention may also be used in liquid forms such as oily suspensions, solutions, emulsions, syrups, together with antioxidants, preservatives, binders, wetting agents, lubricants, thickeners, flavours and aromas. Solid and liquid formulations may be prepared according to generally known methods applicable for preparing pharmaceutical formulations.
Pharmaceutical formulations according to the invention may also be used in the form of suppositories containing usual carriers and other pharmaceutically acceptable auxiliary substances for application in human and veterinary medicine.
Suitable antibiotics which are optionally used in a pharmaceutical formulation according to the invention are not only those antibiotics which are susceptible to degradation by xcex2-lactamases, but also antibiotics having a high degree of resistance to some xcex2-lactamases. As penicillins benzylpenicillin, phenoxymethylpenicillin, ampicillin, amoxycillin, carbencillin etc. may be used and as cefalosporins cefalexin, cefaclor, cefadroxyl, cefpyramid, ceftriaxone, cefixim etc. may be used. When the compounds of the formula I in the form of an acid or of pharmaceutically acceptable salts or esters thereof are combined with another antibiotic in the pharmaceutical formulation, the weight ratio between the compound of the formula I or the pharmaceutically acceptable salt or ester thereof and the antibiotic is in the range from 10:1 to 1:20 and in an amount from 0.1 mg to 1000 mg.
The amount of the compound of the formula I in the form of an acid or of pharmaceutically acceptable salt or ester thereof in pharmaceutical formulations according to the invention is in the range between 0.05 and 100 mg of the active substance per kg of body weight or animal weight, preferably between 0.1 and 40 mg of the active substance per kg of body weight or animal weight. Pharmaceutical formulations according to the invention may be applied from one to four times a day depending upon the way of the application and upon the health condition of the patient.
The invention also relates to novel processes for the preparation of novel compounds of the formula I.
Compounds of the formula I may be prepared in such a way that a compound of the formula II 
wherein R7 represents an easily removable ester protective group such as alkyl, benzyl, 4-nitrobenzyl, tert-butyl group and the ring marked C has the meaning given in the compound of the formula I, is converted to a compound of the formula I in three steps according to novel processes hitherto not disclosed in the literature for tricyclic carbapenems. 
In the first step the hydroxy group of the compound of the formula II is converted into a more easily leaving group, which is better suitable for elimination, in such a way that a compound of the formula III is formed, wherein the radical R8 may be:
an acyl radical from an aliphatic acid, usually aliphatic acid with 1 to 10 carbon atoms, and preferably R8 means acetyl radical,
an alkanesulfonyl radical from an alkanesulfonic acid and usually R8 may mean alkanesulfonyl with 1 to 10 carbon atoms which may be once or several times substituted on carbon atoms, preferably with halo, such as fluoro, and preferably R8 means methanesulfonyl, ethanesulfonyl, fluoromethanesulfonyl, trifluoromethanesulfonyl,
an arenesulfonyl radical from arenesulfonic acid and usually R8 may mean benzenesulfonyl which may be once or several times substituted, preferably in para position (as p-toluenesulfonyl), or naphthalenesulfonyl,
heteroarenesulfonyl radical from heteroarenesulfonic acid and preferably R8 means quinoline-8-sulfonyl.
The conversion of a compound of the formula II to a compound of the formula III may be best carried out in two ways, which, however, does not exclude other ways known in the literature. The reaction is carried out at a temperature from xe2x88x9278xc2x0 C. to the reflux temperature of the solvent, preferably between xe2x88x9220xc2x0 C. and room temperature, and reactants are most frequently added to the reaction mixture at a temperature from xe2x88x9220xc2x0 C. to 0xc2x0 C., and later on the temperature may rise to room temperature. The reaction may be completed immediately after adding the last reactant, in 24 hours at the latest, usually after 10 to 120 minutes.
According to the first method the reaction of a compound of the formula II to a compound of the formula III is carried out by means of a reactive acid derivative in the presence of a base in an inert organic solvent. As the reactive acid derivative reactive derivatives of carboxylic or sulfonic acids may be used such as acid halides R8Xxe2x80x2, wherein Xxe2x80x2 represents halo, preferably chloro, and preferable reactants are methanesulfonyl chloride, p-toluenesulfonyl chloride, acid anhydrides (R8)2O, preferably acetanhydride, trifluoromethane sulfonanhydride. As bases there may be used organic nitrogen bases such as mono, di or trisubstituted alkylamines, preferably triethylamine, diethyl isopropylamine, N,N,Nxe2x80x2,Nxe2x80x2-tetramethylethylene diamine, or amidines such as 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene, or aromatic bases such as pyridine, imidazole, 4-dimethylaminopyridine. As inert organic solvents there may be used inert solvents such as ethers, preferably tetrahydrofuran, dioxan, aromatic hydrocarbons such as benzene, toluene, heterocycles such as pyridine, acetonitrile, and preferably as inert organic solvents chlorinated solvents are used, preferably dichloromethane. As the inert organic solvent also a compound may be used which simultaneously participates in the reaction also in the function of a reactive acid derivative such as compounds N,N-dimethylformamide and N,N-dimethylacetamide.
According to the second method the reaction of a compound of the formula II to a compound of the formula III is carried out by means of the acid R8OH which is bound with condensation agents (reactants for elimination of water) to hydroxy group in 4 position of a compound of the formula III, preferably in the same organic solvents as disclosed in the first method. As condensation agents N,Nxe2x80x2-carbonylimidazole, diphenylphosphorylazide, diphenylphosphorylcyanide, carbodiimides such as dicyclohexylcarbodiimide, N-ethyl-Nxe2x80x2-(3-dimethylaminopropyl)-carbodiimide, diazacarboxylates in the presence of phosphines (reaction according to Mitsunobu, O. Mitsunobu, Synthesis 1981, 1), preferably diethyl diazadicarboxylate in the presence of triphenylphosphine, may be used. Besides this method also other methods suitable for elimination of water may be used. 
In the second step of the conversion of a compound of the formula II to a compound of the formula I, a substituent R8 from a compound of the formula III is cleaved by means of base-catalyzed elimination to obtain a compound of the formula IV. In the reaction as the base there may be used inorganic bases (such as alkali metal carbonates), organic bases such as alkali metal salts of alcohols (such as sodium methoxide, sodium ethoxide, potassium tert-butoxide) and of amides (such as lithium bis(trimethysilyl)amide, lithium diisopropylamide), alkyl metals (such as butyllithium), aryl metals (such as phenyllithium), organic amines (such as triethyl amine, ethyl diisopropylamine, tetramethylethylenediamine), amidines (such as 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene), preferably bases of amidine type are used in the reaction as strong non-nucleophilic bases which are soluble in aprotic organic solvents. As a solvent an inert organic solvent is used such as alcohols (such as methanol, ethanol, tert-butanol, yet not in the case when amides, alkyl metals and aryl metals are used as bases), hydrocarbons such as aromatic hydrocarbons (such as benzene, toluene), ketones (such as acetone), esters (such as ethyl acetate), acetonitrile, amides (such as N,N-dimethylformamide, N,N-dimethylacetamide, hexamethylphosphoric triamide), dimethyl sulfoxide, preferably ethers such as tetrahydrofuran, dioxan, and chlorinated solvents such as dichloromethane, chloroform, dichloroethane. The reaction is carried out at temperatures from xe2x88x9278xc2x0 C. to the reflux temperature of the solvent, preferably at a temperature between xe2x88x9220xc2x0 C. and room temperature. The reaction may be carried out for 5 minutes to 48 hours, mostly for 1 to 6 hours.
The reaction from a compound of the formula II to a compound of the formula IV may be carried out directly without isolating a compound of the formula III. In these cases the group R8O is spontaneously cleaved from the compound of the formula III in the following ways:
during the conversion of the compound of the formula II to the compound of the formula III under the influence of a base used in the reaction with a reactive acid derivative,
thermically during the conversion of the compound of the formula II to the compound of the formula III with a reactive acid derivative or during the process of isolation of the compound of the formula III (elimination takes place here without the presence of a base),
under the influence of the chromatographic carrier (such as silica gel) during the chromatographic purification of the compound of the formula III.
The group R8O from a compound of the formula III is cleaved in such a way that the methyl group in the ethylidene substituent in 6 position of carbapenem ring of a compound of the formula IV is configured around the double bond as (Z) or (E) and in most cases the (E) isomer is prevalent.
In the process for preparing a compound of the formula IV also two diasteroisomers 4-(R) and 4-(S) are formed since in 4 position of the carbapenem ring of the compound of the formula IV representing a joint point with the ring marked C, a new chiral centre is formed. Individual diastereoisomers may be separated by chromatographic purification on a chromatographic carrier.
Both diastereoisomers may be separated already at the compound of the formula II in such a way that the synthesis process from the compound of the formula II over the compound of the formula III to the compound of the formula IV is carried out with already separated isomers. When due to certain reasons (such as poor resolution) diastereisomers of the compound of the formula II cannot be separated, individual diasteroisomers may be obtained from diasteroisomeric mixtures of the compounds of the formula III or IV also in later steps in the same way as disclosed above.
Both geometric isomers are isolated by means of chromatographic purification of the isomeric mixture of the compound of the formula IV on silica gel.
The synthesis routes disclosed for preparing compounds of the formula IV do not exclude the use of other methods known from the literature. 
In the last step of the conversion of a compound of the formula II to a compound of the formula I, from a compound of the formula IV an ester protective group R7 is eliminated from the substituent COOR7 in 2 position of carbapenem ring of the compound of the formula IV according to generally known methods for eliminating a particular protective group. Thus the allyl group is preferably removed with Pd(0) catalysts (preferably tetrakis(triphenylphosphine)palladium (0)) in the presence of phosphine (preferably triphenylphosphine) and under the addition of an alkali metal cation (preferably alkali metal salt of 2-ethylhexanoic acid). Benzyl and 4-nitrobenzyl groups are removed by catalytic hydrogenation in the presence of the alkali metal cation donor (T. W. Green in Protective Groups in Organic Synthesis, John Willey and Sons, New York 1981, p. 191) and as the catalyst preferably palladium on a carrier (such as 10% palladium on active carbon) is used and as the alkali metal cation donor preferably sodium or potassium hydrogen carbonate is used, the product of both kinds of conversions being a compound of the formula I wherein X represents an alkali metal. If the protective group is removed without the presence of a cation, a compound of the formula I in the form of an acid (X represents hydrogen in this case) is isolated and a salt thereof is prepared by neutralization or reprecipitation according to methods known from the literature. The protective group R7 may also be removed in the presence of a nitrogen base such as ammonia, amines (such as trimethylamine, triethylamine, N,Nxe2x80x2-dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, 2-piperidinyl), amidines (such as 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene, 3,3,6,9,9-pentadimethyl-2,10-diazabicyclo[4.4.0]dec-1-ene), guanidines (such as guanidine, cyanoguanidine) or other nitrogen bases (such as 4-dimethylaminopyridine, imidazole) so that in the obtained compound of the formula I X represents the above mentioned protonated nitrogen bases.
When in the ring marked C of the compound of the formula IV there are substituents protected with well-known protective groups, these substituents should, before step 3, be first removed by well-known deprotection methods except in cases when these protective groups are removed with the reactants mentioned in the description of step 3.
The compounds of the formula I in the form of an acid or a salt (X represents an alkali metal) may be converted by methods well-known in the Prior Art to biologically acceptable esters of the formula I (in this case X represents an organic allyl radical, most frequently of the type CHR5OCOR6).
The disclosed synthesis routes for the conversion of the compound of the formula IV to the compound of the formula I do not exclude the use of other methods described in the literature.
The starting compound of the formula II is prepared from (3R,4R)-4-acetoxy-3-((R)-1-(tert-butyldimethylsilyloxy)ethyl)azetidine-2-one which is commercially available as Azetidon(copyright)xe2x80x94Kaneka (Japan) according to known processes disclosed in the literature:
T. Murayama et al., Tetrahedron Lett. 35, (1994) 2271; J. Tsuji, Tetrahedron 42 (1986) 4401; P. J. Reider et al., Tetrahedron Lett. 23, (1982) 379; A. Yoshida et al., Tetrahedron Lett. 25, (1984) 2793; I. Ernest et al., J. Am. Chem. Soc. A0, (1979) 6310; A. Afonso et al., J. Am. Chem. Soc. 104, (1982) 6139; R. Di Fabio et al., Bioorg. Med. Chem. Lett. 5, (1995) 1235; F. A. Corey and R. J. Sundberg, Advanced Organic Chemistry Second Edition, Plenum Press New York and London, 1983, Annual Reports in Organic Synthesis 1975-1989, Academic Press Inc., San Diego; T. W. Green, Protective Groups in Organic Synthesis, John Willey and Sons, New York 1981, p. 45.
The invention is disclosed in detail by the following Examples which in no way should be construed as limitative thereto. Silica gel of the company Merck, trade mark Kieselgel 60, particle size 0.063-0.200, in a 50 to 100 fold amount of the input substance was used for column chromatography and the course of the reactions and preparative chromatography was followed by thin layer chromatography on glas plates of the company Merck, trade mark Kieselgel 60 GF254.